Conventionally, a cylindrical container, called a column, having a liquid inlet and outlet in the center of an upper and lower circular plates, is filled with a particulate filler, called a gel, having a particle diameter of ten to several hundred microns, so that removal, separation and purification of a substance are performed utilizing the interaction between solute molecules and the gel when a liquid is made to flow from either the upper or lower liquid inlet or outlet by a pump or the like.
Incidentally, treatment steps using a column requires a large amount of transfer liquid (transfer phase) since a solution comprising a target material to be separated and purified is passed while contacting with a filler in the column in one direction. As a result, the materials included in the solution are separated into the materials to be captured by the filler and the materials to be flown out with the transfer liquid. Where a useful target material is included in the materials that are flown out, there was a problem that the target material to be separated and purified is diluted since the transfer liquid is in a large amount.
For example, where a target useful protein is separated from a residual liquid from which major proteins have been removed, as in the treatment of the proteins in the blood, there was a problem that the residual liquid is diluted since a large amount of transfer liquid is flown in the column in the treatment using a conventional column, and concentration of the residual liquid is required, and thus the treatment becomes complex.
Accordingly, the inventors of the present application have made a pipette tip comprising an attachment opening that is to be attached to a nozzle for sucking and discharging a gas or to a connecting member attachable to the nozzle, and an opening that allows flow-in and flow-out of a liquid in response to the suction and discharging of the gas to adsorb or capture the biological material in the liquid, and enclosed a support that may react with or bond to the biological material in the pipette tip, so that the direction of the flow of the liquid contacting with the support becomes bidirectional. This constitution allows separation of a target material using relatively a small amount of a liquid by repeating suction and discharging with respect to the pipette tip. The inventors have made possible to perform separation and purification more efficiently and rapidly than the treatment using a conventional column, and aimed at improving the separation performance of the target material using an easier and smaller scale structure (Patent Document 3).
However, there were problems that where air bubbles are included between gel particles during enclosing a support such as a support for affinity chromatography in a column or a pipette tip, passage of the liquid is obstructed and unreacted parts may be generated, and thus the column or pipette tip needs to be stored in the state that the air bubbles between the gel particles are completely removed.
Furthermore, there were problems that where the gel enclosed in the column or pipette tip is dried, the volume is changed, and as a result, the performance of the column may be deteriorated since the column correlates with the reaction volume, and that cracking may occur in the gel and the gel is disabled.
Moreover, even where the gel is fixed on thread or beads, there was a problem that air bubbles may not be removed after drying, and deformation or separation may occur due to drying.
In addition, although proteins, enzymes, antibodies and the like may be solid-phased in dried state and stored, those forming a subunit such as metal proteins and flavin having a solid-phase factor had a problem that they are dissociated once they are dried and may not be reconstructed.
Moreover, it was necessary to store proteins and the like in an antiseptic agent such as sodium azide so as to prevent decomposition.    Non Patent Document 1: “Liquid Chromatography Q&amp; A” (published by Gihodo Shuppan Co., Ltd., June 2006, written by Itaru Matsushita)    Non Patent Document 2: “Reality of Liquid Chromatography” (published by Sankyo Publishing Co. Ltd., 1976, written by Akira Etoh)    Patent Document 1: WO2006/1073170